- Methods in Cell Biology, Volume 75: Cytometry 4th Edition. New Developments. (2004) Darzynkiewick, Roederer and Tanke (eds)
- Current Protocols in Flow Cytometry
- Course Notebook - 27th Annual Course in Flow Cytometry
These flow protocols have been validated by members of the Flow Core.
Flow Core Database - fill out a sort form or report an issue here
Sort Form Information - information about filling in the sort form
The reference bead that is used in the facility is from Spherotech. We use the Rainbow Calibration Particles, 6th peak of 8. The part number for this product is RCP-30-5A-6 and the current lot that we use is AC01. With the recent change of bead lots, we also are supplying this excel file to illustrate the changes between the previous bead lot (AB02) and the new lot (AC01).
Double click the "shortcut to transfer" icon on the desktop of the instrument computer. An explorer window with a folder for each PI will open. NOTE - you will be able to view the contents of the folders but will not be able to read the contents of any of these folders, even your own. These folders are write only from the instruments. As before, drag the folders you want to transfer from the FCS folder to your group folder.
To recover your data on your lab computer:
Click "Start" then in the "Search programs and files" field type:
\\smdnas\FCC_Transfer\PILastName_FirstInitial (ie. bushnell_t)
If you've had Windows remember your credentials then you may have to delete
those. You can do this by clicking the "Start" button and then in the
"Search programs and files" field that appears type "credential manager".
You should see a suggested icon of a safe appear in the list above the
search field that says "Credential Manager". Click this, it will open a
new window showing your saved credentials. Under "Windows Credentials"
delete anything related to RDS01. If you are in doubt feel free to delete
a Windows Credential, the worst case scenario is you will have to re-input
your password for something else.
Click "Start" and then "Run".In that field type
\\smdnas\FCC_Transfer\PILastName_FirstInitial (ie. bushnell_t)
Click "Go" in Finder, choose "Connect to Server" then type:
smb://smdnas/fcc_transfer/PILastName_FirstInitial (ie. bushnell_t)
smb://smdnas.urmc-sh.rochester.edu/fcc_transfer/PILastName_FirstInitial (ie. bushnell_t)
If you are prompted for a username and password use your URMC
network credentials (what you would use for webmail or to log onto a
Windows computer here).
Data will be automatically removed from the transfer partition after two weeks.
This is done similarly to the Transfer space. Here we have shared documents that are available to all registered users. This includes past lectures, IDEAS software, and other goodies.
Start Menu > Run...
Finder >Go > Connect to server
PDF available (behind the firewall)
The URMC Flow Cytometry Core facility (FCC), under the direction of Timothy Bushnell, Ph.D., is a full service facility with approximately 1100 sq. feet of newly renovated lab space, employing a staff of 6. The facility currently has a range of digital flow cytometers including a 2-laser, 4-color Accuri C6, a 3-laser, 8-color BD-FACSCanto-II, two 3-laser, 12-color BD LSR-II’s, and two 4-laser, 18-color BD LSR-II’s. Additionally, the facility is equipped with a 3-laser, 13-color FACSAria-Iiu, a 4-laser, 18-color FACSAria-II, both of which are equipped with aerosol management systems, automatic cell deposition units, and are housed within Baker Bio-Protect-III biosafety hoods. The facility also has a 3-laser, 6-channel IS100 and 4-laser, 12-channel ImagestreamX imaging flow cytometers from Amnis. The facility has three dedicated analysis workstations for investigators with standard flow cytometric analysis software (Flowjo, IDEAS, Modfit). All data generated in the FCC is archived onto a 10-terabyte server with daily tape backup.
The FCC has established a series robust QA/QC protocols and procedures that are designed to ensure the instruments are performing optimally. These include standard quality control measurements, as recommended by the instrument manufacturers. Additional detailed instrumentation characterization and QC are performed based on best practices in the field. All QA/QC information is available to investigators and instrumentation characterization data is available on the FCC’s website.
All investigators go through a robust training program to allow them run the analyzers independently, allowing for experienced users to have 24/7 access. All booking is done via an on-line calendar system. All sorting is done by the FCC staff, and is available to investigators from ~10 am till 9:30 pm daily.
- Enos, M., Bancos, S., Bushnell, T., and Crispe, I.N. (2008) E2F4 modulates differentiation and gene expression in hematopoietic progenitor cells during commitment to the lymphoid lineage J Immunol 180:3699-707
- McGrath, K.E., Kingsley, P.D., Koniski, A..D., Porter, R.L., Bushnell, T.P., and Palis, J.(2007) Enucleation of primitive erythroid cells generates a transient population of "pyrenocytes" in the mammalian fetus. Blood 111:2409-17
- Guzman, M.L., Li, X., Corbett, C.A., Rossi, R.M., Bushnell, T., Liesveld J.L., Herbet, J., Young, F., and Jordan C.T. (2007) Rapid and selective death of leukemia stem and progenitor cells induced by the compound 4-benzyl, 2-methyl, 1,2,4-thiadiazolidine, 3,5 dione (TDZD-8). Blood 110:4436-44
- Ryan, E.P., Bushnell, T.P., Friedman, A.E., Rahman, I., and Phipps, R.P. (2007) Cyclooxygenase-2 selective inhibition reduces B lymphoma survival by inducing oxidative stress and decreasing intracellular glutathione Cancer Immunol Immunother 57:347-58
- Neering, S.J., Bushnell, T., Sozer, S., Ashton, J., Rossi, R.M., Wang, P.-Y., Bell, D.R., Heinrich, D., Bottaro, A, and Jordan C.T. (2007) Leukemia stem cells in a genetically defined murine model of blast crisis CML. Blood 110:2578-85.
- Graf, B., Bushnell, T., and Miller , J. (2007) LFA-1-mediated T cell costimulation through increased localization of TCR/class II complexes to the cSMAC and exclusion of CD45 from the immunological synapse. J Immunol., 179:1616-24
- Neff-LaFord, H., Teske, S., Bushnell, T.P., and Lawrence, B.P. (2007) Aryl hydrocarbon receptor activation during influenza virus infection unveils a novel pathway of IFN-a production by phagocytic cells J Immunol 179:247-55
- Kingsley, PD, Malik J, Emerson RL, Bushnell TP, McGrath KE, Bloedorn LA, Bulger M, and Palis J (2006) “Maturational” globin switching in primary primitive erythroid cells. Blood 107:1665-1672
Location/Time - Rm 3-4103 @ 4PM (in the flow core hallway)
Schedule: Sept 3rd*not a Monday, 15th and 29th.
next lecture coming soon!
Comments about the Flow Core?